反义VEGF硫代寡脱氧核苷酸抑制裸鼠人肝癌移植瘤生长的研究

Inhibition of Growth of Human Hepatocellular Carcinoma in Nude Mice Model by Antisense Phosophorothioate Oligodeoxynuclieotidesin

目的 :检测硫代修饰的反义脱氧寡核苷酸 (ASPODN)抑制人肝癌血管形成和生长的作用 ,以探讨ASPODN的临床应用前景。方法 :传代培养的肝癌细胞系HepG2 ,接种于裸鼠颈背部皮下 ,自 4 8h始局部注射ASPODN和等量的PBS ,彩色多普勒能量图 (CDE)检测移植瘤内新生血管的数目和分布 ;LSAB法免疫组化染色检测移植瘤中微血管密度 (MVD) ;FCM分析裸鼠移植瘤细胞周期动力学和凋亡率。结果 :局部注射ASPODN组和PBS对照组 ,裸鼠人肝癌移植瘤瘤重及MVD平均值分别为 ( 1.2 8±0 .2 6)g、2 6.90± 7.4 2和 ( 3.4 6± 0 .86)g、50 .36± 10 .2 7(P <0 .0 1) ;局部注射ASPODN组 ,用CDE动态检测到移植瘤内的血管数目及血流丰富程度明显低于PBS组P <0 .0 1;经ASPODN治疗后的裸鼠人肝癌移植瘤细胞凋亡率 ( 7.4 8± 1.64) %高于PBS对照组 ( 2 .75± 0 .2 2 ) % (P <0 .0 1)。结论 :VEGFAS PODN明量抑制肝癌血管形成和生长 。

Objective To study the inhibition effect of antisense phosophorothioate oligodeoxynuclieotidesin(ASPODN) on growth and angiogensis of human hepatocellular carcinoma in nude mice model,investigate whether ASPODN of VEGF can become kind of drug for antitumor gene therapy.Methods 5×10 6/ml HepG 2 were inoculated subcutaneously to 12 nude mices,they were randomized into 2 groups.From the 2 nd day,ASPODN and PBS were given.The number and distribution of angiogenesis of sucutaneous tumors were detected with color doppler energy (CDE),microvesse density was dete cted with immunohistochemical stain.A cytokinetic study and apoptosis rate of human hepatocellular carcinoma in nude mice model was performed by FCM.Results The tumor weight and MVD of ASPODN group were(1.28±0.26)g and 28.90±7.42;PBS control group were (3.46±0.86)g and 50.36±10.28,respectively( P <0.01).The angiogenesis numers and distribution of human hepatocellular carcinoma in nude mice model in CDE was correlation with MVD( P <0.05).Apotosis index in ASPODN group7.48±1.64 was higher than that in PBS group 2.75±0.22 ( P <0.01).Conclusions The growth and angiogenesis of human hepatocellular carcinoma in nude mice model can be inhibited by ASPODN;VEGF ASPODN may become an new antitumor angiogenesisi gene therapy drugs.